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Magnetic cell sorting for enriching Schwann cells from adult mouse peripheral nerves

2003; Elsevier BV; Volume: 123; Issue: 2 Linguagem: Inglês

10.1016/s0165-0270(02)00349-7

1872-678X

Jan Manent, Ksénia Oguievetskaia, Jan Bayer, Nancy Ratner, Marco Giovannini,

Neuroblastoma Research and Treatments

We have devised a simple method to purify mitotically active Schwann cells (SC) from peripheral nerves of adult mice. Nerves were predegenerated in vitro for 7 days and after dissociation cells were plated on poly-l-lysine/laminin coated dishes in N2 serum-free culture medium supplemented with forskolin and heregulin-β1. Primary cultures were purified from contaminating fibroblasts by magnetic cell sorting (MACS) based on SC membrane specific expression of p75NGFR and enriched to about 99% of SC after MACS from 34 to 91% before sorting. After sorting, purified adult mouse SC were propagated for three passages until confluent to a total surface of 160 cm2 per mouse (two sciatic and two trigeminal nerves). In addition, we show that this method can be used to purify tumoral SC from mouse NF2-related schwannomas.